Method for inducing menses

ABSTRACT

THERE IS DISCLOSED HEREIN A METHOD FOR INDUCING MENSES IN PREGNANT PRIMATES WHEREIN CHORIONIC GONADOTROPIN OR PREGNANT MARES&#39;&#39; SERUM GONADOTROPIN, OR MIXTURES THEREOF, ARE ADMINISTERED AS A SINGLE DOSE OR AS CLOSELY SPACED DIVIDED DOSE DURING THE PERIOD OF TIME FROM DAY 14 TO DAY 35 OF GESTATION.

United States Patent 3,816,617 METHOD FOR INDUCING MENSES Upendra K.Banik, Pierrefonds, Quebec, Canada, assignor tls YAmerican Home ProductsCorporation, New York, No Drawing. Filed July 25, 1972, Ser. No. 274,960Int. Cl. A61k 17/00, 17/06 US. Cl. 424-100 1 Claim ABSTRACT OF THEDISCLOSURE There is disclosed herein a method for inducing menses inpregnant primates wherein chorionic gonadotropin or pregnant mares serumgonadotropin, or mixtures thereof, are administered as a single dose oras a closely spaced divided dose during the period of time from day 14to day 35 of gestation.

This invention relates to therapeutic compositions containinggonadotropins and to a method for using such compositions for inducingmenstruation or menses.

More specifically, this invention resides in the concept ofadministering to primates, for example, humans, monkeys or baboons,during the period of time from about day 14 to about day 35 ofgestation, an effective dose of a composition to induce menstruationcomprising, for example, chorionic gonadotropin or pregnant mares serumgonadotropin.

In another aspect, this invention has relativeness to todays concern forovercoming medical and biosocial problems associated with our increasingpopulation. Today, more and more couples are turning to means forachieving a planned family. For a woman of childbearing age theoccurrence of menstruation has always been the most convincing andassuring sign that an unplanned pregnancy has not taken place. Thispresent invention provides means whereby the desired menstruation andaccompanying peace of mind may be obtained without the use of mechanicalor therapeutic contraceptives, which are so often relied upon today.Furthermore, the method of this invention does not require theadministration of prolonged or continuous regimens such as associatedwith todays therapeutic contraceptives, thereby avoiding the hazards oflong-term therapy.

I have found that menstruation may be induced in primates from about 14to about day 35 of gestation by administering thereto a menses inducingdose of chorionic gonadotropin or pregnant mares serum gonadotropin, ormixtures thereof. For the purpose of this disclosure, day 0 of gestationis defined as the 14th day after the beginning of the last menses forhumans, and as the day of mating for other primates.

Chorionic gonadotropin and pregnant mares serum gonadotropin have beenfound to be suitable to be administered according to the method of thisinvention. It has been established that these gonadotropins may beadministered safely to primates; for example, see I. H. Leathem and A.E. Rakoff, Amer. J. Obstet. GynecoL, 56, 521 (1948) and referencestherein, and E. B. Astwood in The Pharmacological Basis of Therapeutics,4th ed., L. S. Goodman and A. Gilman, eds., The Macmillan Co., New York,1970, pp. 1512-1537. The preparation and characterization of thesegonadotropins have been described; see for example, Gonadotropins:Physiochemical and Immunological Properties, Ciba Foundation Study GroupNo. 22, G. E. W. Wolstenholme and 1. Knight, eds., J. & A. ChurchillLtd., London, 1965. Therapeutically they are used primarily for thetreatment of infertility and cryptorchidism. In addition thesegonadotropins have been suggested for alleviating a variety of otherconditions related to hypogonadal dysfunction, such pr I ICC ashypoovarianism, amenorrhea, functional uterine bleeding, threatened andhabitual abortion, as well as for the treatment of testicularhypofunction such as hypogenitalism or hypogonadotropic eunuchoidism.

More specifically, several investigators have established the essentialrole played by chorionic gonadotropin in primates in maintaining thecorpus luteum especially during the first eight to ten weeks ofpregnancy; see, I. T. Bradbury er al., Recent Progr. Hormone Res., 5,151 (1950), C. A. Gemzell et al., J. Clin. Endocrinol. Metab., 18, 133(1958 and J. D. Neill et al., Endocrinol., 84 45 (1969). Moreover, asimilar role has been indicated for pregnant mares serum gonadotropinduring pregnancy in the horse, E. B. Astwood, cited above. Accordinglythe use of exogenously administered gonadotropins has been studiedextensively in women and found in certain cases to stimulate andmaintain the integrity of the ovary and corpus luteum; for example, seeE. C. Hamblen, Endocrinol., 24, 848 (1939) and R. B. Greenblatt inOfiice Endocrinology, Charles C. Thomas, Springfield, Ill., 1944, pp.79-86.

These investigations of the last forty years have definitely establishedthat the gonadotropins, especially Chorionic gonadotropins, are capableof maintaining and prolonging the functional life span of the corpusluteum in non-gravid humans [for example, see F. W. Hanson et al., J.Clin. Endocrinol., 32, 211 (1971)] and nongravid monkeys [F. L. I-Iisaw,Yale J. Biol. Med, 17, 119 (19441945)]. These latter findings concerningthe life span of the corpus luteum led several investigators to proposeon a seemingly logical basis that chorionic gonadotropin or pregnantmares serum gonadotropin therapy is indicated for the treatment andprevention of threatened abortion; for example, see L. W. Mason, Amer.J. Obstet. Gynec., 35, 559 (1938), W. E. Brown and J. T. Bradbury, Amer.J. Obstet. Gynec., 53, 749 (1947), P. H. Fried and A. E. Rakoff, J.Clin. Endocrinol. Metabol., 12, 321 (1952); see also S. L. Isreal inMenstrual Disorders and Sterility, 5th ed., Harper & Row, New York,1967, p. 604, who states that, in pregnancy, chorionic gondotropin maybe of value as a stimulant to the corpus luteum until the placentaperforms its luteal function It is therefore well established in humansand monkeys that the physiological and therapeuticf attribute ofchorionic gonadotropin is to be luteotropic and that of pregnant maresserum gonadotropin to be promotion of functional follicles, although afew studies in rodents have shown that these gonadotropins may interferwith pregnancy, see R. M. Coco, Amer. J. Physiol, 137, 143 (1942) and W.H. Yang and M. C. Chang, Endocrinol., 83, 217 (1968).

It is well known, however, that because of the unpredictable nature ofapplying the findings respecting the corpus luteum of one species toanother and because of the fundamental physiologic differences in thereproductive processes of rodents and primates, these later findings inrats, rabbits and hamsters have not lead any one to expect thatgonadotropins could be used as menses inducers in primates". Thisunpredictability respecting species has been expressed by severalauthorities in this field, see e.g. Greenblatt or Brown and Bradbury,both cited above. B. V. Caldwell et al., J. Reprod. Fert., Suppl. 8, 59(1969) has expressed it in the following way: Throughout the classMammalia there is a wide variation in corpora lutea life-span andfunction, and the effects of operations such as hysterectomy andhypophysectomy in various species are by no means uniform.

It will readily be seen from the foregoing review of the literature thatit was the consensus of scientific opinion that gonadotropins wereuseful in re-establishing normal gonadal function in primates in casesof gonadal hypofunction, and that gonadotropins played an important rolein maintaining a normally functioning corpus luteun which is essentialto protect and preserve the integrity of the fetus during earlypregnancy.

In marked contradistinction to those opinions I have now found that theadministration of gonadotropins to primates during early pregnancy,i.e., from about day 14 to about day 35 of gestation, induces menseswithin 3-7 days after administration. In the light of the foregoing itwill be readily understood, therefore, that my discovery of the hereindisclosed menses inducing properties of gonadotropins in primates wasindeed paradoxical, unexpected and surprising.

When used in accordance with this invention, the gonadotropins areadministered intramuscularly, intravenously, subcutaneously, orintravaginally to the pregnant primate, preferably with a pharmaceuticalcarrier in dosage units.

For intramuscular, intravenous or subcutaneous, administration,commercial preparations of these gonadotropins, for example, seeRemingtons Pharmaceutical Sciences, 14th ed., Mack Publishing Co.,Easton, P'a., pp. 956-957, are injected into the primate. Morespecifically, in accordance with the method of this invention, dosesranging from 200-1200 i.u./kg, of chorionic gonadotropin or -120i.u./kg. of pregnant mares serum gonadotropin, or appropriatelyproportioned mixtures thereof, are administered intramuscularly orsubcutaneously either as a single dose or in divided doses over aclosely spaced period of time, for example, one to three days, toprimates within the period of time of from about day 14 to about day 35of gestation. For convenience, it is preferred to administer this dosageas a single dose. Thereafter, menses commence usually within three toseven days. When the gonadotropins of this invention are administeredintravenously, the same dosages for the gonadotropins are used,administered preferably at a continuous rate over a period of 30 minutesto three hours.

For vaginal applications similar doses of the gonadotropins of thisinvention are prepared with suitable carriers to form tablets, capsules,creams or the like suitable for insertion into the vagina. For example,a suitable tablet or capsule may be prepared using lactose, starch orsucrose as the carrier. Other suitable carriers and methods forcompounding them into tablets, capsules, creams and the like aredescribed in Remingtons Pharmaceutical Sciences, cited above.

The effects of administering the gonadotropins according to the methodof this invention was specifically demonstrated in the following manner.

Female rhesus monkeys weighing from 2.5 to 6.0 kg. and having a regularmenstrual cycle of between 25 to 30 days were housed (one per cage)during the 11th to 15th day of their cycle with a male rhesus monkey ofproven fertility. The female monkeys were observed for mating signs,i.e., the presence of spermatozoa in the vagina. When mating signs wereobserved the female monkey was placed in a separate cage.

Thereafter pregnancy was established in the female monkey by gestationalbleeding occurring approximately 22 to 30 days after the last menses andby the detection of serum or urinary chorionic gonadotropin by a methodusing the bioassay of H. O. Burdick et al., EndocrinoL, 33, 1 (1943). Asin this particular method, a 24 hour collection of urine was treatedwith twice its volume of acetone. The resulting precipitate was driedunder reduced pressure and dissolved in distilled water. The solutionwas injected intraperitoneally into adult diestrus mice at 3 pm. Withrespect to the serum, a sample of it was diluted two times withdistilled water and injected into the test animals as before. Theanimals were sacrificed on the following morning. The observation thatfreshly shed ova were present in the fallopian tubes of the miceindicated that chorionic gonadotropin was present in the original urineor serum sample, thereby indicating pregnancy. [Other pregnancy testsmay also be used; see for example, B. M. Hibbard, -Brit. Med. 1., 1, 593(1971).] When pregnancy was diagnosed, the day on which mating signswere observed was assigned day 0 of gestation. The diagnosis ofpregnancy was verified further by rectal palpation during the second andthird week of gestation. The pregnant monkeys were then divided into twogroups of six monkeys. The first group of monkeys were given a singledose of either 5000 i.u. of human chorionic gonadotropin or 1,000 i.u.of pregnant mares serum gonadotropin reconstituted with a sterilediluent. This dose was administered between day 20 and day 35 ofgestation. The second group served as control and received only aninjection of the sterile diluent.

Within seven days after the injection, a menstrual type of bleedingoccurred in more than 50% of the monkeys in the first or treated group;whereas bleeding was found to occur in less than 10% of the second orcontrol group. Practically every monkey in which bleeding or menses hadthereby been induced in the first group returned subsequently to aregular cycling period.

I claim:

1. A method for inducing menses in pregnant primates selected from theclass consisting of monkeys and humans which comprises administering tosaid primates a menses inducing dose of a material, selected from thegroup consisting of chorionic gonadotropin in doses of from 200- 1200i.u./kg, pregnant mares serum gonadotropin in doses of from 10-120i.u./kg, and a mixture thereof, within the period of time of from aboutday 14 to day 35 of gestation to produce menstruation and a return tothe normal menstrual cycle.

References Cited Greenwald et al.: Chem. Abst., vol. 71 (1969), p.19231p.

Moor et al.: Chem. Abst., vol. 71 (1969), p. 5720*5r.

SAM ROSEN, Primary Examiner US. Cl. X.R. 42410l, 108

